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Greening disease is the most destructive disease of citrus in many countries in the worl. It in very serve in Vietnam. One of the most powerful tool to diagnostic this disease is PCR. However, it is very expensive and complex, not easy for provincal centers to diagnostic. The experiment were carried out to improve the protocol with the aim to reduce the cost of testing disease and also to reduce the risk to the environment due to reducing a lot of toxic chemicals. The results are included:
Use centrifuge at room temperature instead of use cool centrifugate were not effect to PCR results with the
Primers OI I/OI 2 c. In the DNA extraction steps, the chemical were reduced from 750µl of Cloroform: isopropanol to 550µl isopropanol from 650µl to 450µl and TE 1X form 100µl to 70µl, the PCR result were 100%. The primer OI 1/OI 2 c concentration was reduced from 0.5µl to 0.2µl the PCR result were 100%. The clearness of the band were the same at all concntration of this primers. The concentration of Taq DNA polymerase were reduced from 1.25U to 0.6U but PCR results were 100%. The PCR Kit to diagnostic Greening diseases were gaven good results after one month produced (stored at 4oC). Concentration of Tween 20 at 0.05% and 0.03% were given good rusults (PCR is 1005) (store at 4oC).
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National Center for Scientific and Technology Information
